EP1754534A1 - Polymère hydrophile réticulé - Google Patents

Polymère hydrophile réticulé Download PDF

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Publication number
EP1754534A1
EP1754534A1 EP05016846A EP05016846A EP1754534A1 EP 1754534 A1 EP1754534 A1 EP 1754534A1 EP 05016846 A EP05016846 A EP 05016846A EP 05016846 A EP05016846 A EP 05016846A EP 1754534 A1 EP1754534 A1 EP 1754534A1
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EP
European Patent Office
Prior art keywords
alkyl
aryl
radical
independently
alkoxy
Prior art date
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EP05016846A
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German (de)
English (en)
Inventor
Matthias Dr. Jöhnck
Eckhard Dr. Sabrowski
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Merck Patent GmbH
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Merck Patent GmbH
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Application filed by Merck Patent GmbH filed Critical Merck Patent GmbH
Priority to EP05016846A priority Critical patent/EP1754534A1/fr
Priority to CN201110266314.2A priority patent/CN102417557B/zh
Priority to CA2617645A priority patent/CA2617645C/fr
Priority to PCT/EP2006/005484 priority patent/WO2007014591A1/fr
Priority to US11/997,613 priority patent/US7951885B2/en
Priority to ES06754224.1T priority patent/ES2528426T3/es
Priority to CN2006800285993A priority patent/CN101258175B/zh
Priority to EP06754224.1A priority patent/EP1910433B1/fr
Priority to JP2008524371A priority patent/JP5475284B2/ja
Priority to KR1020087005296A priority patent/KR101289911B1/ko
Publication of EP1754534A1 publication Critical patent/EP1754534A1/fr
Priority to US12/947,900 priority patent/US8765897B2/en
Priority to US14/173,218 priority patent/US20140155565A1/en
Withdrawn legal-status Critical Current

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J20/282—Porous sorbents
    • B01J20/285—Porous sorbents based on polymers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F216/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal or ketal radical
    • C08F216/12—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal or ketal radical by an ether radical
    • C08F216/14—Monomers containing only one unsaturated aliphatic radical
    • C08F216/1416—Monomers containing oxygen in addition to the ether oxygen, e.g. allyl glycidyl ether
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F222/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical and containing at least one other carboxyl radical in the molecule; Salts, anhydrides, esters, amides, imides, or nitriles thereof
    • C08F222/36—Amides or imides
    • C08F222/38—Amides
    • C08F222/385—Monomers containing two or more (meth)acrylamide groups, e.g. N,N'-methylenebisacrylamide

Definitions

  • the present invention relates to a hydrophilic crosslinked polymer, preferably in the form of porous particles, as well as its preparation and use.
  • the polymer according to the invention is produced by polymerization from chain-forming hydrophilic vinyl ethers and crosslinking N, N'-divinyl compounds, preferably heterocyclic N, N'-divinyl compounds.
  • WO 03/104294 discloses polyvinyl ethers, in particular for use in solid phase synthesis.
  • the disclosed polymers usually show a very strong swelling behavior.
  • WO 9513861 discloses polyvinyl ethers for separation purposes which are prepared by cationic polymerization. The described cationic polymerization must be carried out in combination with a complex protective group chemistry.
  • EP 0 482 339 discloses a copolymer based on allyl monomers. Especially on EP 0 482 339 based copolymer of 1,2, -dihydroxy-3-allyloxy-propane and N, N'-methylenebisacrylamide, commercially available under the name FractoPrep® (Merck KGaA, Germany), shows in terms of hydrophilicity, hydrolytic stability and pressure-flow stability for the Biochromatography good properties. Disadvantage of this material, however, is the complicated production process by means of inverse suspension polymerization.
  • the object of the present invention was therefore to provide a hydrophilic polymeric carrier material which fulfills the aforementioned requirements with respect to porosity, hydrophilicity, stability to hydrolysis and pressure-flow stability and is also easy to prepare.
  • hydrophilic polymers with good properties in particular for biochromatography, can be prepared by copolymerization of hydrophilically substituted alkyl vinyl ethers with crosslinking, preferably heterocyclic N, N'-divinyl compounds.
  • the polymers are preferably prepared by suspension polymerization.
  • R4 in formula I is an alkyl radical, a cycloaliphatic radical or an aryl radical bearing at least one hydroxyl group.
  • R4 is in formula I. a straight-chain or branched C 1 to C 10 alkyl radical in which one or more nonadjacent methylene groups may be replaced by O, S, SO 2 or NH and / or in which one or more H atoms are independently of one another C 1 -C 6 -alkyl, C 5 C10-aryl, C1-C6-alkoxy or C1-C6-alkyl-OH and in which at least one OH group is present either on the C1 to C10 alkyl radical or on a substituent, or a cycloaliphatic radical having typically 5 to 10 C atoms, in which one or more non-adjacent methylene groups may be replaced by O, S, SO 2 or NH and / or in which one or more H atoms of the cycloaliphatic radical are independent of each other C1-C6-alkyl, C5-C10-aryl, C1-C6-alkoxy or C1-
  • the hydrophilically substituted alkyl vinyl ether used is 1,2-ethanediol monovinyl ether, 1,3-propanediol monovinyl ether, 1,4-butanediol monovinyl ether, 1,6-hexanediol monovinyl ether or diethylene glycol monovinyl ether and cycloaliphatic vinyl ether as cyclohexanedimethanol monovinyl ether.
  • the crosslinker used is divinylethyleneurea (1,3-divinylimidazolin-2-one) or divinylpropyleneurea (1,3-divinyltetrahydropyrimidin-2-one).
  • the polymer is porous with pore sizes between 2 and 200 nm.
  • the polymer is in the form of particles having a diameter between 3 and 300 microns.
  • the polymer is derivatized with separation effectors.
  • the polymer is derivatized with tentacle-like structures which have been applied to the polymer by graft polymerization under cerium (IV) catalysis.
  • the tentacle-like structures are copolymers made from two or more different monomers.
  • a free-radical suspension polymerization is carried out.
  • a free-radical suspension polymerization is carried out in water as suspending agent in the presence of at least one suspension stabilizer and optional further additives, preferably inorganic salts and surface-active compounds.
  • the temperature during the polymerization is between 40 and 100 ° C.
  • hydrophilic substituted alkyl vinyl ethers and crosslinkers are used in a mass ratio of between 10:90 and 80:20.
  • the present invention also provides a chromatography column, capillary or cartridge containing a polymer according to the invention as sorbent or carrier material.
  • the present invention also relates to the use of a hydrophilic crosslinked polymer according to the invention as a sorbent in chromatography, as a carrier material for the immobilization of biologically or catalytically active substances or as a carrier material for a solid phase synthesis.
  • Figure 1 shows the retention behavior of the polymer according to the invention when giving up proteins. Further details can be found in Example 2.
  • FIG. 1 shows the experimentally determined distribution coefficients Kd of dextrans with different molecular weights. Further details can be found in Example 3.
  • FIG. 3 shows the storage stability of a polymer according to the invention in dilute sodium hydroxide solution. Further details can be found in Example 4.
  • cycloaliphatic radical means a saturated hydrocarbon radical in which all or at least part of the hydrocarbon units are part of a cyclic structure.
  • N (C 1 -C 8) alkyl 2 denotes a nitrogen which is at least substituted by two alkyl radicals.
  • the two alkyl radicals can independently of one another have 1 to 8 C atoms in straight-chain or branched form.
  • a heteroaryl radical is an at least partially aromatic radical which differs from an aryl radical in that one or more nonadjacent C atoms are replaced by N, S or O. It is clear to the person skilled in the art that, owing to the valences, the heteroatoms may optionally be substituted by H, C1-C6-alkyl or C1-C6-alkyl-OH.
  • the polymer is formed by copolymerization of at least one compound from the group of the hydrophilically substituted alkyl vinyl ethers according to formula I and at least one compound from the group of crosslinkers according to formula II and / or III and / or IV.
  • Preferred is only one compound from the Group of hydrophilically substituted alkyl vinyl ethers according to formula I and a compound selected from the group of crosslinkers of formula II, III or IV.
  • One or more compounds from the group of hyrophil-substituted alkyl vinyl ethers of the formula I and / or one or more compounds from the group of crosslinkers corresponding to formula II and / or III and / or IV can be used.
  • the polymerization mixture further polymerizable compounds may be added, which in the Polymer skeleton are polymerized.
  • these are compounds having at least one polymerizable double bond.
  • only one compound from the group of hydrophilically substituted alkyl vinyl ethers according to formula I and a compound from the group of crosslinkers according to formula II, III or IV is used.
  • the hydrophilically substituted alkyl vinyl ether is 1,2-ethanediol monovinyl ether, 1,3-propanediol monovinyl ether, 1,4-butanediol monovinyl ether, 1,6-hexanediol monovinyl ether or diethylene glycol monovinyl ether and used as cycloaliphatic vinyl ether Cyclohexandimethanolmonovinylether.
  • Crosslinkers used are preferably compounds of the formula II. It is preferred to use divinylpropyleneurea (1,3-divinyltetrahydropyrimidin-2-one) or, with particular preference, divinylethyleneurea (1,3-divinylimidazolin-2-one).
  • the proportion of the hydrophilically substituted alkyl vinyl ethers in the weight of the polymer is typically between 1 (weight)% and 90 (weight)% or a maximum weight fraction of the alkyl vinyl ether, which corresponds to a molar ratio of 2: 1 based on a bifunctional crosslinker, if the alkyl vinyl ether is not homopolymerized ,
  • the proportion of hydrophilically substituted alkyl vinyl ethers is preferably between 10 and 80%, particularly preferably between 35 and 60%.
  • the proportion of the crosslinker is between 10 and 99, preferably between 20 and 90%, particularly preferably between 40 and 65%.
  • a radical polymerization is carried out.
  • a through Radical initiated suspension polymerization also called bead polymerization.
  • water-insoluble or poorly soluble monomers which are also referred to as oil
  • an oil-in-water suspension normal phase
  • water-soluble monomers are dissolved in water and suspended in an organic solvent which is immiscible or only slightly water-miscible and polymerized (inverse polymerization).
  • the suspension polymerization is carried out in a known manner.
  • the suspending agent water is typically used in the normal phase suspension polymerization. It is known to add one or more stabilizers and / or one or more surfactant compounds to the suspending agent.
  • macromolecular stabilizers are polyvinylpyrolidone, polyacrylates, polycarboxylates, polyacrylamide, polyvinyl alcohol, hydroxyalkylcellulose, methylcellulose or polyethylene glycols.
  • inorganic compounds z.
  • calcium phosphate or magnesium hydroxide can be used as stabilizers.
  • aqueous phase buffer substances such. B. Na 2 HPO 4 and NaH 2 PO 4 are added.
  • Suitable surface-active compounds are, in particular anionic and nonionic surfactants, such as ethoxylates of long-chain alcohols, ethoxylated mono-, di- and trialkylphenols, alkali metal and ammonium salts of C 12 - C 18 alkyl sulfonic acids or aryl sulfonic acids.
  • anionic and nonionic surfactants such as ethoxylates of long-chain alcohols, ethoxylated mono-, di- and trialkylphenols, alkali metal and ammonium salts of C 12 - C 18 alkyl sulfonic acids or aryl sulfonic acids.
  • the organic phase also called the oil phase
  • the oil phase contains, in addition to the monomers, inert solvents (porogens) in order to set the desired pore sizes.
  • Suitable organic solvents are z.
  • aliphatic hydrocarbons C 6 - C 18
  • cycloaliphatic compounds such as cyclohexane, aromatic hydrocarbons such as toluene, ethylbenzenes or xylenes or alcohols such as aliphatic C 4 - C 12 alcohols z.
  • heptanol dodecanol, cyclohexanol, polyethylene glycols or polypropylene glycols with different molecular weight or esters of aliphatic carboxylic acids such as butyl acetate or propyl propionate or esters of aromatic carboxylic acids such as propyl benzoate or esters such as butyl glycol acetate or glycerol triacetate or ethers such as di-n-butyl ether, di-n-amyl ether, diphenyl ether , Ethylene glycol monophenyl ether and ethylene glycol diethyl ether.
  • solvents can either act as swelling or precipitating agents for the polymers, thus affecting porosity.
  • swelling agents in the polymers according to the invention are lower alcohols, aliphatic esters or aromatic hydrocarbons such as toluene.
  • precipitants are aliphatic hydrocarbons such as dodecane or gasoline mixtures.
  • free-radical initiators are typically added to the organic phase.
  • free-radical initiators are organic peroxides, such as di-tert-butyl peroxide, dibenzoyl peroxide, bis (o-methylbenzoyl) peroxide, tert-butyl hydroperoxide, cumene hydroperoxide, di-iso-propyl peroxide dicarbonate, cyclohexanone peroxide, or aliphatic azo compounds, such as ⁇ , ⁇ '-azodiisobutyronitrile , Azobis-cyanovaleric acid, 1,1'-azo-cyclohexane-1,1'-dicarboxylic acid nitrile or azodicarbonamide. Preference is given to ⁇ , ⁇ '-azodiisobutyronitrile.
  • activatable initiators can be used according to the invention as radical initiators.
  • phase ratios in the suspension polymerization can typically be varied in a volume ratio of between 1:20 (oil phase to aqueous phase) and 2: 1; ratios between 1: 5 and 1: 1 are preferred, more preferably between 1: 3 and 1: 1.
  • the ratio of organic solvent to monomer is typically between 1:20 and 4: 1, preferably between 1: 5 and 3: 1, more preferably between 1: 1 and 2.5: 1.
  • the concentration of the initiator based on the amount of monomer is typically between 0.01 (weight)% and 10 (weight)%, preferably between 0.1 and 7 (weight)%, and more preferably between 0.2 and 7 (weight)%.
  • the concentration of stabilizers based on the amount of suspending agent, usually water, is between 0.01 and 3 (weight)%, typically between 0.02 and 2 (weight)%, more preferably between 0.05 and 0.5 (weight)%.
  • the suspension polymerization is then carried out by mixing the two phases with vigorous stirring. In addition, typically for a period of 1 to 20 hours to 20 to 100 ° C, preferably 40 to 100 ° C tempered. Typically, the mixture is heated to 40 to 100 ° C over a period of 0.5 to 5 hours and then postpolymerized again at 70 to 100 ° C for several hours. During the whole time should be stirred.
  • These temperatures refer to a system in which water is used as a suspending agent. When using other polar solvents instead of water, depending on the boiling point of the solvent other temperature ranges may be possible or useful.
  • the polymer can be brought into the desired particle size distribution by sieving or another type of classification.
  • the polymerization according to the invention can be carried out continuously.
  • static mixers in particular micromixers or tracked mixers, are particularly preferably used for particle generation.
  • the spatial distribution of two components to be mixed is influenced by the flow guidance, eg by separating and reuniting, twisting, distorting and widening, so that large interfaces are created for the diffusive exchange between the components and a high shear forces particularly uniform particle size distribution can be achieved.
  • Various mixers and their construction are known. For example, such systems are described in EP 1177243 ,
  • the micromixer continuously flows through the aqueous phase with a defined volume flow. Subsequently, the oil phase is fed via the second feed, also with a defined, precisely adjusted volume flow.
  • the ratio of the two volume flows can be in a wide range and depends on the desired particle size, the type and distribution of the particle sizes, the viscosity of the aqueous and the oil phase depending on the reaction temperature and the reaction kinetics and the required ratio of the porogens in the final mixture Achieving a suitable pore structure.
  • the volume flow ratio between the aqueous phase and the oil phase is preferably 1:10 to 100: 1, in particular 2: 1 to 20: 1.
  • the suspension leaving the static mixer is polymerized in a continuous reactor.
  • particle diameters between 3 .mu.m and 500 .mu.m, preferably between 3 .mu.m and 300 .mu.m, can be produced by means of suspension polymerization, depending on the type of agitator and the rotational speed.
  • the particles may be irregularly shaped or, preferably, spherical.
  • the polymers of the invention are prepared by emulsion polymerization, e.g. Particle diameter between 20 nm and 3000 nm, preferably between 100 and 500 nm are generated.
  • the polymers of the invention are preferably present as non-porous or preferably porous particles.
  • the pore sizes can be between 2 and 300 nm, depending on the type and amount of the porogenic solvent used. Pore sizes between 2 and 200 nm are preferred.
  • the polymers can also be present in the form of membranes, fibers, hollow fibers, coating or as a monolithic shaped body.
  • Monolithic shaped bodies are three-dimensional bodies, for example in cylindrical form. These shaped bodies preferably have a mono- or bimodal pore structure.
  • the polymers according to the invention can be used as composite materials, ie, for example, as a coating of, for example, inorganic particles or shaped bodies, or im Mixture with eg inorganic components.
  • An example of this are particles of the polymer according to the invention, which are themselves magnetizable by copolymerizing magnetizable particles or a magnetizable core.
  • hydrophilic, crosslinked polymers according to the invention are particularly suitable as sorbents in chromatography, as support materials for the immobilization of biologically and / or catalytically active substances or as support materials for solid phase syntheses of e.g. Biopolymers such as nucleic acids or peptides or for combinatorial chemistry.
  • the materials of the invention are characterized by a good hydrolytic stability, especially in basic medium, and by a good pressure-flow stability. Furthermore, due to their high hydrophilicity, they are particularly suitable for biochromatographic processes.
  • the examples additionally provide experimental data on these properties.
  • the polymers according to the invention are particularly suitable as sorbents. For this purpose, they are filled in a known manner in chromatography columns or capillaries. Likewise, the polymer of the invention may be packed in cartridges for chromatographic or other purposes. The present invention therefore also chromatographic columns, capillaries or cartridges containing a polymer according to the invention as a sorbent or carrier material.
  • the polymers according to the invention can be used in their native form, ie without further derivatization steps, or can be provided with other or additional functionalities by one or more derivatization steps. In particular, they can be derivatized with separation effectors.
  • the covalent attachment of the separation effectors is usually via the functional groups present on the polymer, e.g. Hydroxyl groups, e.g. to form an ester or preferably an ether function, directly or via a linker or spacer.
  • the linkage with the base material via a cerium (IV) -catalyzed graft polymerization to form a C-C linkage with the base material.
  • Thiophilic radicals are for example in EP 0 165 912 disclosed.
  • the polymer may e.g. to introduce epoxy groups with glycidyl compounds such as butanediol diglycidyl ether.
  • the polymer according to the invention can be provided as a base material by graft polymerization with tentacle-like structures, which in turn can carry the corresponding separation effectors or can be functionalized with them.
  • the grafting is preferred according to EP 0 337 144 carried out.
  • the generated chain is linear and linked to the base material via a monomer unit.
  • the base material according to the invention is suspended in a solution of monomers, preferably in an aqueous solution.
  • the grafting of the polymeric material is effected in the course of a conventional redox polymerization with exclusion of oxygen.
  • cerium (IV) ions are used because this catalyst forms radical sites on the surface of the base material from which the graft polymerization of the monomers is started.
  • the polymerization is terminated by termination reactions involving the cerium salts. Therefore, the (average) chain length is determined by the concentration ratios of the base material, the initiator and the Monomers influenced. Furthermore, uniform monomers or mixtures of different monomers can be used; in the latter case, grafted copolymers are formed.
  • Suitable monomers for the preparation of the graft polymers are monomers corresponding to formula V or VI.
  • CR * R ** CR 11 -Y (formula V)
  • the product obtained is preferably then converted into a separating material having hydroxyl groups.
  • This conversion into a hydroxyl phase is achieved by a known mild alkaline or acid saponification.
  • the reaction with methanolic K 2 CO 3 solution at room temperature described for example by Y. Tezuka et al., in Macromol. Chem. 186 , 685-694 (1985).
  • R 11 is preferably H, ie the acrylic acid derivatives are preferred.
  • Y in formula V is preferably , -COCHR 15 R 16 or -CH 2 NH 2 , second preferred -CN or -CHO.
  • R 15 and R 16 independently of one another denote H or an alkyl group having up to 5 C atoms. Preferably, at least one of R 15 and R 16 is H.
  • the following radicals are particularly preferred: acetyloxy, propionyloxy, butyryloxy, valeryloxy and hexanoyloxy radicals.
  • Z in formula V denotes -OR 14 , -OH or -NR 12 R 13 , preferably -NR 12 R 13 . Compounds in which Z - NR 12 R 13 and one of the radicals R 12 and R 13 is H are preferred.
  • the radicals R 12 and / or R 13 are preferably an alkyl, phenyl, phenylalkyl or alkylphenyl group, it being possible for the alkyl and / or the phenyl group to be monosubstituted or polysubstituted, preferably monosubstituted or disubstituted, particularly preferably monosubstituted by an alkoxy, cyano, amino, mono- or dialkylamino, trialkylammonium, carboxyl, sulfonic acid, acetoxy or acetamino radical.
  • the radicals R 12 and / or R 13 are preferably alkyl, alkoxyalkyl, cyanoalkyl, aminoalkyl, mono- or dialkylaminoalkyl, trialkylammoniumalkyl, carboxyalkyl or sulfonic acid alkyl having up to 10 C atoms, preferably up to 6 C atoms, particularly preferably up to 4 C atoms in the alkyl group, which may be linear or branched.
  • R 12 and / or R 13 are preferably methyl, ethyl, propyl, butyl, pentyl, hexyl, methoxymethyl, ethoxymethyl, 2-methoxyethyl, 2-, 3- or 4-oxapentyl, 2-, 3-, 4- or 5 Oxahexyl, 2-, 3-, 4-, 5- or 6-oxaheptyl, Isopropyl, 2-butyl, isobutyl, 2-methylbutyl, isopentyl, 2-methylpentyl, 3-methylpentyl, 2-oxa-3-methylbutyl, 3-oxa-4-methylbutyl, 2-methyl-3-oxapentyl, 2-methyl 3-oxahexyl, and also heptyl, octyl, nonyl or decyl.
  • R 12 and / or R 13 are preferably cyanomethyl, cyanoethyl, cyanopropyl, cyanobutyl, cyanopentyl, cyanohexyl, 2-cyanopropyl, 2-cyanobutyl, carboxylmethyl, carboxylethyl, carboxylpropyl, carboxylisopropyl, carboxylbutyl, carboxylpentyl, carboxylhexyl, carboxyl-2-methylpropyl, Carboxyl-2-methylbutyl, sulfonic acid methyl, sulfonic acid ethyl, sulfonic acid propyl, sulfonic acid butyl, sulfonylpentyl, sulfonic acid hexyl, sulfonic acid 2-methylpropyl, sulfonic acid 2-methylpropyl, sulfonic acid 2-methylpropyl, sulfonic acid 2-methylpropyl, s
  • the alkyl groups are preferably monosubstituted by an amino, mono- or dialkylamino or trialkylammonium group.
  • the alkyl groups may be identical or different and have up to 10, preferably up to 6 C atoms, more preferably up to 4 C atoms, and therefore preferably mean dimethylaminoethyl, diethylaminoethyl, methylaminoethyl, methylaminopropyl, dimethylaminopropyl, ethylaminoethyl, propylaminoethyl, propylaminopropyl , Dipropylaminoethyl, dipropylaminobutyl, diethylaminoethyl, trimethylammoniumethyl, trimethylammoniumpropyl, Trimethylammoniumbutyl, triethylammoniumethyl, triethylammoniumpropyl, triethylammoniumethyl, aminoethyl,
  • R 12 and / or R 13 also has the meaning of a phenyl group, which is preferably monosubstituted by cyano, cyanoalkyl, amino, aminoalkyl, mono- or dialkylamino, alkyl, alkoxy, alkoxyalkyl, mono- or dialkylaminoalkyl, trialkylammonium or trialkylammoniumalkyl, Carboxy, carboxyalkyl, sulfonic acid or sulfonic acid alkyl.
  • the preferred meanings of these substituents correspond to the preferred alkyl groups and substituted alkyl groups given above.
  • the substituent on the phenyl group is preferably in the p position.
  • R 12 and / or R 13 are also preferred for R 12 and / or R 13 .
  • R 12 and / or R 13 is an alkylphenyl or phenylalkyl group, wherein the preferred meanings given for the alkyl, substituted alkyl or substituted phenyl groups are also to apply.
  • substituted phenyl groups are for example particularly preferred: 4-cyanophenyl, 4-alkylphenyl, 4- (N, N-dimethylamino) -phenyl, 4- (N, N-dialkylaminoethyl) -phenyl, 4-ethoxyphenyl, 4-ethoxyethylphenyl, 4-trialkylammonium phenyl, 4-carboxylphenyl, 4-sulfonic acid phenyl, phenylethyl, 4- (N-ethylamino) phenylpropyl or 4-cyanophenylethyl.
  • R 12 and / or R 13 is a cyclic or bicyclic radical which may be aromatic or saturated, having 5-10 C atoms, in which one or more CH- or CH 2 - Groups by N or NH, N or NH and S, or N or NH and O are replaced mean.
  • R 12 and / or R 13 are preferably also a pyridine radical, imidazolyl radical, indolyl radical, furthermore preferably a pyrrole, pyrimidine, pyrazine, quinoline or isoquinoline radical.
  • R 12 and / or R 13 may also be a thiazole, thiadiazole, morpholine, triazine, piperazine, benzothiazole, purine, pyrazole, triazole, pyrrolidine or isoxazole radical.
  • Particularly preferred are the aromatic, heterocyclic radicals.
  • the radicals R 12 and R 13 must, in order to arrive at suitable exchangers, be coordinated so that either both radicals contain an acidic or basic group or one of the radicals is neutral. It is not difficult for a person skilled in the art to assign the groups accordingly and thus to put together suitable radicals for R 12 and R 13 , depending on the function and function of the desired ion exchanger.
  • one of the two radicals R 12 and R 13 is a neutral radical.
  • R 14 is preferably alkyl, alkoxyalkyl, cyanoalkyl, carboxyalkyl or sulfonic acid alkyl having up to 10 C atoms, preferably having up to 6 C atoms, more preferably having up to 4 C atoms in the alkyl group, which may be linear or branched.
  • R 14 therefore preferably denotes methyl, ethyl, propyl, butyl, pentyl, hexyl, methoxymethyl, ethoxymethyl, 2-methoxyethyl, 2-, 3- or 4-oxapentyl, isopropyl, 2-butyl, isobutyl, 2-methylbutyl, isopentyl, 2 Methylpentyl, 3-methylpentyl, 2-oxa-3-methylbutyl, 3-oxa-4-methylbutyl, 2-methyl-3-oxapentyl or 2-methyl-3-oxahexyl. Also preferred are alkyl groups which are substituted by a cyano, carboxy or sulfonic acid group.
  • R 14 is preferably cyanomethyl, cyanoethyl, cyanopropyl, cyanobutyl, cyanopentyl, cyanohexyl, 2-cyanopropyl, 2-cyanobutyl, carboxylmethyl, carboxylethyl, carboxylpropyl, carboxylisopropyl, carboxylbutyl, carboxylpentyl, carboxylhexyl, carboxyl-2-methylpropyl, carboxyl-2-methylbutyl , Sulfonic acid methyl, sulfonic acid ethyl, sulfonic acid propyl, sulfonic acid butyl, sulfonylpentyl, sulfonic acid hexyl, sulfonic acid 2-methylpropyl, sulfonic acid 2-methylbutyl, sulfonic acid 3-methylbutyl, Sulfonic acid 2-methylpentyl, sulfonic acid 3-methylbut
  • R 14 also has the meaning of a phenyl group which is preferably monosubstituted by cyano, cyanoalkyl, alkyl, alkoxy, alkoxyalkyl, carboxy, carboxyalkyl, sulfonic acid or sulfonic acid alkyl.
  • the preferred meanings of these substituents correspond to the preferred alkyl groups and substituted alkyl groups given above.
  • the substituent on the phenyl group is preferably in the p position.
  • R * and R ** in the monomers of the formula V are preferably H.
  • R * and R 11 in formula VI are preferably H.
  • chains are grafted onto the base material which have between 2 and 100, preferably 5 and 60, in particular between 10 and 30 monomer units.
  • sorbents in which only one type of separation effector is present in the base matrix
  • co-polymers or "mixed-mode” sorbents in which there are at least two different types of separation effectors in the sorbent.
  • "single mode" sorbents separate the solutes based on a type of interaction between separation effector and solute.
  • solute separation relies on different types of interaction between separation effector and solute.
  • Such chromatographic methods are known as mixed-mode chromatography. In the simplest case, one uses, for example, the hydrophobicity of the basic matrix, in order to introduce an ionic Separation effector provide a suitable for mixed-mode chromatography sorbent.
  • mixed-mode sorbents are accessible.
  • the disadvantage of both approaches is that the steric relationship between the different types of separation effectors is undefined. This disadvantage is avoided in mixed-mode sorbents, which combine the different separation effectors in one ligand. This ligand is bound to the base matrix.
  • Both of the aforementioned types of mixed-mode sorbents are of LW McLaughlin (1989) in Chem. Rev. 89, 309-319 described in a review article.
  • a mixed-mode sorbent the various Separationss capableoren bound in a single ligand, can be provided from a chloropropyl substituted carrier material by conversion of the chlorine derivatives in bromine derivatives and subsequent reaction with benzyldimethylamine:
  • the ligand is bound via a C3 chain to the base matrix. This is followed by a dimethyl-substituted ammonium group to which a lipophilic benzyl radical is attached.
  • a reverse arrangement of hydrophobic and ionic separation effectors in a ligand is obtainable by reacting aminopropyl substituted substrate with p-diethylaminobenzoic acid: in this ligand, the tertiary amino group is oriented distally of the base matrix, while in the former variant the hydrophobic benzyl group is located distally.
  • Further embodiments of such ligands with two separation effectors are disclosed in the following references: WO 96/09116 . WO 97/29825 .
  • EP 04028798.9 become more complex ligands with more than one separation effector, as well as anion exchange groups, whose nitrogen as Azaderivat of multi-ring structures, such as Chininuklidinsystem, is present. Examples for this are:
  • a jacketed reactor 1050 ml of deionized water (deionized water) are introduced and dissolved therein 0.32 g of tributyl phosphate, 4.2 g of sodium sulfate, 0.01 g of sodium laurylsulfonate and 2.1 g of polyvinyl alcohol as a stabilizer.
  • the organic phase consisting of 96.25 g of butanediol monovinyl ether, 78.45 g of divinylethyleneurea, 175 g of dodecanol and 4.2 g of ⁇ , ⁇ '-azo-diisobutyronitrile, is added after mixing and dissolving the ingredients at room temperature to the aqueous phase with stirring.
  • the suspension is stirred vigorously and then heated to 70.degree. After about 2 hours, the polymerization is continued at 80 ° C. for 4 hours. After cooling to room temperature, the polymer is filtered off with suction, the porogen is removed by steam distillation and the product is stored in 20% ethanolic solution. There are obtained 130 g of polymer, based on dry matter. The average grain size is 80 ⁇ m.
  • the swelling volume of the reaction product is 4.3 ml / g.
  • the determination of the retention behavior of proteins as a function of the salt concentration is used to investigate the protein binding behavior of the unmodified carrier material. Since proteins bind only nonspecifically on the unmodified carrier material, it is advantageous if the least possible background binding occurs.
  • the binding behavior of the material according to the invention is compared to the unmodified copolymer of 1,2-dihydroxy-3-allylaxy-propane and methylenebisacrylamide, available commercially under the name FractoPrep® (Merck KGaA, Germany).
  • Figure 1 shows the binding behavior of the proteins lysozyme and bovine serum albumin as well as NaNO 3 .
  • the abbreviation FP stands for FractoPrep®
  • B / D stands for the inventive copolymer of butanediol monovinyl ether and divinylethyleneurea.
  • the recovery of the measured proteins expressed as the quotient of the photometrically determined amount of protein eluted from the column under the running conditions to the protein applied to the column, is greater than 99% under all experimental conditions.
  • An exemplary pore structure of the polymer according to the invention is shown by way of example in FIG. 2 by the experimentally determined distribution coefficients Kd of dextrans with different molecular weights, expressed by the viscosity radius of the dextrans.
  • Eluent 100 mM NaCl, 20 mM NaH 2 PO 4 , pH 7.2
  • FIG. 3 shows the results. It can be clearly seen that storage in dilute sodium hydroxide solution has no effect on the distribution coefficient.

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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
EP05016846A 2005-08-03 2005-08-03 Polymère hydrophile réticulé Withdrawn EP1754534A1 (fr)

Priority Applications (12)

Application Number Priority Date Filing Date Title
EP05016846A EP1754534A1 (fr) 2005-08-03 2005-08-03 Polymère hydrophile réticulé
ES06754224.1T ES2528426T3 (es) 2005-08-03 2006-06-08 Polímero hidrófilo reticulado
CA2617645A CA2617645C (fr) 2005-08-03 2006-06-08 Polymere reticule hydrophile
PCT/EP2006/005484 WO2007014591A1 (fr) 2005-08-03 2006-06-08 Polymere reticule hydrophile
US11/997,613 US7951885B2 (en) 2005-08-03 2006-06-08 Hydrophilic crosslinked polymer
CN201110266314.2A CN102417557B (zh) 2005-08-03 2006-06-08 亲水交联聚合物
CN2006800285993A CN101258175B (zh) 2005-08-03 2006-06-08 亲水交联聚合物
EP06754224.1A EP1910433B1 (fr) 2005-08-03 2006-06-08 Polymère hydrophile réticulé
JP2008524371A JP5475284B2 (ja) 2005-08-03 2006-06-08 親水性架橋ポリマー
KR1020087005296A KR101289911B1 (ko) 2005-08-03 2006-06-08 친수성 가교 중합체
US12/947,900 US8765897B2 (en) 2005-08-03 2010-11-17 Hydrophilic crosslinked polymer
US14/173,218 US20140155565A1 (en) 2005-08-03 2014-02-05 Hydrophilic crosslinked polymer

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EP05016846A EP1754534A1 (fr) 2005-08-03 2005-08-03 Polymère hydrophile réticulé

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114007737A (zh) * 2019-07-03 2022-02-01 默克专利股份公司 抗体药物缀合物纯化

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Publication number Priority date Publication date Assignee Title
DE3344912A1 (de) 1983-12-13 1985-06-20 Hoechst Ag, 6230 Frankfurt Vernetzte polymerisate, verfahren zu ihrer herstellung und ihre anwendung
EP0165912A2 (fr) 1984-05-17 1985-12-27 Jerker Porath Adsorbant à base de thioéther pour la séparation de protéines et analogues
EP0266503A1 (fr) * 1986-08-28 1988-05-11 Hoechst Aktiengesellschaft Polymères réticulés et leur procédé de préparation
DE3714276A1 (de) 1987-04-29 1988-11-17 Hoechst Ag Hydrophile, vernetzte polymerisate, verfahren zu ihrer herstellung und ihre verwendung
EP0337144A1 (fr) 1988-03-31 1989-10-18 MERCK PATENT GmbH Matériaux de séparation
EP0482339A1 (fr) 1990-09-15 1992-04-29 Röhm Gmbh Support hydrophile, activable, sous forme de perles et hautement réticulé
WO1995013861A1 (fr) 1993-11-17 1995-05-26 Pharmacia Biotech Ab Procede de separation et polymeres de synthese pouvant etre utilises comme milieux de separation dans ce procede
WO1996009116A1 (fr) 1994-09-23 1996-03-28 Massey University Resines chromatographiques et leurs procedes de mise en ×uvre
US5599702A (en) * 1990-12-24 1997-02-04 Hoechst Aktiengesellschaft D-amino acid oxidase from Trigonopsis variabilis immobilized on porous copolymer beads
WO1997029825A1 (fr) 1996-02-19 1997-08-21 Amersham Pharmacia Biotech Aktiebolag Procede de separation chromatographique de peptides et d'acide nucleique et nouvelle matrice d'echange d'ions a haute affinite
WO2000069872A2 (fr) 1999-05-14 2000-11-23 Promega Corporation MATRICE ECHANGEUSE D'IONS DEPENDANT DU pH, UTILISEE POUR ISOLER DES ACIDES NUCLEIQUES
WO2001038228A1 (fr) 1999-11-22 2001-05-31 Amersham Biosciences Ab Procedes d'adsorption d'echange anionique et echangeurs anioniques a liaison thioether
EP1177243A1 (fr) 1999-05-06 2002-02-06 MERCK PATENT GmbH Procede de fabrication de polymeres en perles
WO2003104294A1 (fr) 2002-06-05 2003-12-18 Imperial College Innovations Limited Polyvinyle ethers
EP1693108A1 (fr) 2004-12-04 2006-08-23 MERCK PATENT GmbH Matériau de séparation à base d'échangeur d'anions de mode mixte

Patent Citations (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3344912A1 (de) 1983-12-13 1985-06-20 Hoechst Ag, 6230 Frankfurt Vernetzte polymerisate, verfahren zu ihrer herstellung und ihre anwendung
EP0165912A2 (fr) 1984-05-17 1985-12-27 Jerker Porath Adsorbant à base de thioéther pour la séparation de protéines et analogues
EP0266503A1 (fr) * 1986-08-28 1988-05-11 Hoechst Aktiengesellschaft Polymères réticulés et leur procédé de préparation
DE3714276A1 (de) 1987-04-29 1988-11-17 Hoechst Ag Hydrophile, vernetzte polymerisate, verfahren zu ihrer herstellung und ihre verwendung
EP0337144A1 (fr) 1988-03-31 1989-10-18 MERCK PATENT GmbH Matériaux de séparation
EP0482339A1 (fr) 1990-09-15 1992-04-29 Röhm Gmbh Support hydrophile, activable, sous forme de perles et hautement réticulé
US5599702A (en) * 1990-12-24 1997-02-04 Hoechst Aktiengesellschaft D-amino acid oxidase from Trigonopsis variabilis immobilized on porous copolymer beads
WO1995013861A1 (fr) 1993-11-17 1995-05-26 Pharmacia Biotech Ab Procede de separation et polymeres de synthese pouvant etre utilises comme milieux de separation dans ce procede
WO1996009116A1 (fr) 1994-09-23 1996-03-28 Massey University Resines chromatographiques et leurs procedes de mise en ×uvre
WO1997029825A1 (fr) 1996-02-19 1997-08-21 Amersham Pharmacia Biotech Aktiebolag Procede de separation chromatographique de peptides et d'acide nucleique et nouvelle matrice d'echange d'ions a haute affinite
EP1177243A1 (fr) 1999-05-06 2002-02-06 MERCK PATENT GmbH Procede de fabrication de polymeres en perles
WO2000069872A2 (fr) 1999-05-14 2000-11-23 Promega Corporation MATRICE ECHANGEUSE D'IONS DEPENDANT DU pH, UTILISEE POUR ISOLER DES ACIDES NUCLEIQUES
WO2001038228A1 (fr) 1999-11-22 2001-05-31 Amersham Biosciences Ab Procedes d'adsorption d'echange anionique et echangeurs anioniques a liaison thioether
WO2003104294A1 (fr) 2002-06-05 2003-12-18 Imperial College Innovations Limited Polyvinyle ethers
EP1693108A1 (fr) 2004-12-04 2006-08-23 MERCK PATENT GmbH Matériau de séparation à base d'échangeur d'anions de mode mixte

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
L.W. MCLAUGHLIN, CHEM.REV., vol. 89, 1989, pages 309 - 319

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114007737A (zh) * 2019-07-03 2022-02-01 默克专利股份公司 抗体药物缀合物纯化

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